EP2399595A1 - Composition immunopotentiatrice et son procédé de production - Google Patents

Composition immunopotentiatrice et son procédé de production Download PDF

Info

Publication number
EP2399595A1
EP2399595A1 EP10743592A EP10743592A EP2399595A1 EP 2399595 A1 EP2399595 A1 EP 2399595A1 EP 10743592 A EP10743592 A EP 10743592A EP 10743592 A EP10743592 A EP 10743592A EP 2399595 A1 EP2399595 A1 EP 2399595A1
Authority
EP
European Patent Office
Prior art keywords
cells
immune
mre
enhancing composition
cancer
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Granted
Application number
EP10743592A
Other languages
German (de)
English (en)
Other versions
EP2399595A4 (fr
EP2399595B1 (fr
Inventor
Kaoru Mitarai
Yoji Nagahama
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Meisho Co Ltd
Original Assignee
Meisho Co Ltd
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Meisho Co Ltd filed Critical Meisho Co Ltd
Publication of EP2399595A1 publication Critical patent/EP2399595A1/fr
Publication of EP2399595A4 publication Critical patent/EP2399595A4/fr
Application granted granted Critical
Publication of EP2399595B1 publication Critical patent/EP2399595B1/fr
Active legal-status Critical Current
Anticipated expiration legal-status Critical

Links

Images

Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
    • A61K35/66—Microorganisms or materials therefrom
    • A61K35/74—Bacteria
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23K—FODDER
    • A23K10/00—Animal feeding-stuffs
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
    • A23L33/135—Bacteria or derivatives thereof, e.g. probiotics
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K39/02—Bacterial antigens
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K39/02—Bacterial antigens
    • A61K39/07—Bacillus
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K39/39—Medicinal preparations containing antigens or antibodies characterised by the immunostimulating additives, e.g. chemical adjuvants
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/0012—Galenical forms characterised by the site of application
    • A61K9/0019—Injectable compositions; Intramuscular, intravenous, arterial, subcutaneous administration; Compositions to be administered through the skin in an invasive manner
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/0012—Galenical forms characterised by the site of application
    • A61K9/0053—Mouth and digestive tract, i.e. intraoral and peroral administration
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P11/00—Drugs for disorders of the respiratory system
    • A61P11/02—Nasal agents, e.g. decongestants
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P17/00—Drugs for dermatological disorders
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P17/00—Drugs for dermatological disorders
    • A61P17/02—Drugs for dermatological disorders for treating wounds, ulcers, burns, scars, keloids, or the like
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P17/00—Drugs for dermatological disorders
    • A61P17/10—Anti-acne agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P19/00—Drugs for skeletal disorders
    • A61P19/02—Drugs for skeletal disorders for joint disorders, e.g. arthritis, arthrosis
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P19/00—Drugs for skeletal disorders
    • A61P19/08—Drugs for skeletal disorders for bone diseases, e.g. rachitism, Paget's disease
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P27/00—Drugs for disorders of the senses
    • A61P27/02—Ophthalmic agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P3/00—Drugs for disorders of the metabolism
    • A61P3/08—Drugs for disorders of the metabolism for glucose homeostasis
    • A61P3/10—Drugs for disorders of the metabolism for glucose homeostasis for hyperglycaemia, e.g. antidiabetics
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/04—Antibacterial agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/12—Antivirals
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/12—Antivirals
    • A61P31/14—Antivirals for RNA viruses
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/12—Antivirals
    • A61P31/14—Antivirals for RNA viruses
    • A61P31/18—Antivirals for RNA viruses for HIV
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/12—Antivirals
    • A61P31/20—Antivirals for DNA viruses
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P37/00—Drugs for immunological or allergic disorders
    • A61P37/02—Immunomodulators
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P37/00—Drugs for immunological or allergic disorders
    • A61P37/02—Immunomodulators
    • A61P37/04—Immunostimulants
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P37/00—Drugs for immunological or allergic disorders
    • A61P37/08—Antiallergic agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P43/00—Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P9/00—Drugs for disorders of the cardiovascular system
    • A61P9/12—Antihypertensives
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2200/00—Function of food ingredients
    • A23V2200/30—Foods, ingredients or supplements having a functional effect on health
    • A23V2200/324—Foods, ingredients or supplements having a functional effect on health having an effect on the immune system
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K2039/555—Medicinal preparations containing antigens or antibodies characterised by a specific combination antigen/adjuvant
    • A61K2039/55588—Adjuvants of undefined constitution
    • A61K2039/55594—Adjuvants of undefined constitution from bacteria
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K2039/57—Medicinal preparations containing antigens or antibodies characterised by the type of response, e.g. Th1, Th2
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
    • Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
    • Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change

Definitions

  • the present invention relates to a composition that stimulates innate immunity thereby enhancing the antibacterial, antiviral, anti-inflammatory, and anticancer effects and that also affects the lymphocyte immune system so as to suppress inflammation.
  • the composition in accordance with the present invention stimulates innate immune receptors such as TLR, NLR, and RIG, followed by activating innate immune response cells including macrophages, natural killer cells, and, natural killer T cells, as well as activating the lymphocytic immunity interlocked with innate immunity such as Th17 and Th1, or Treg or the like.
  • said composition induces activation of a series of immune processes for producing IL- 21 that suppresses excessive immune responses, thereby enhancing anticancer, antibacterial, antiviral, and anti-inflammatory effects and mitigating autoimmunity due to a runaway of CTL.
  • Innate immunity receptors or similar sensors thereof are widely present in animals, plants, and microorganisms.
  • TLR Toll-like-receptor
  • NLR Nod-like-receptor
  • RLR RLR
  • Ligands are produced in multiples depending on the types of bacteria and viruses, generally, varying in substance and concentration that are generated, resulting in an identification and an innate immunity response, with a subtly different pattern recognition dictated by a combination of their ligand groups.
  • Innate immune response cells such as epithelial cells in humans, have TLR TLR -1 & 2, TLR -6 & 2, TLR-4 & MD-2, and TLR- 5 act on the cell membrane; and bacterial or viral nucleic acids (DNA and /or RNA) degradation fragments from degeneration by lysosomal enzymes in the endosome or phagocytic phagosome activate TLR -3, TLR-7, TLR-8, and TLR-9.
  • NLR receptors that sense low molecular degradation products of bacteria such as NOD1 NOD2, NALP3, and NAIP5, IPAF, and RIG; and MDA-5, and the like that sense viral low molecular degradation products and/or nucleic acids.
  • interleukins and/or type I interferons IFN- ⁇ family, IFN- ⁇ and IFN- ⁇
  • IFN- ⁇ family, IFN- ⁇ and IFN- ⁇ interleukins and/or type I interferons
  • the cells simultaneously release antibacterials such as defensin, cathelicidine, and dermicidin and/or a group of antiviral substances, as many as several hundred types, called the ISG, thereby defending a group of friendly cells from the foreign invaders.
  • antibacterials such as defensin, cathelicidine, and dermicidin and/or a group of antiviral substances, as many as several hundred types, called the ISG
  • the second role of the innate immune receptors is to activate phagocytes present in a multicellular organism with a gut.
  • Phagocytes have developed as cells with a shared responsibility of the moving innate immunity, in which its role is played in humans by macrophages, neutrophils, or the like. Macrophages particularly play important roles as commanders of the innate immune system. When receptors, innate immunity sensors, are stimulated, macrophages, neutrophils, and their peers are activated, launching a framework to fight off invaders such as bacteria and viruses.
  • Vertebrates except for lampreys and hagfish, have developed a lymphocytic immune system that identifies self from non-self, reinforcing the immune system.
  • the mammalian lymphoid immune system has undergone a sophistication and enhancement on the basis of a complement system. On the other hand, this has resulted in patients suffering from allergic and autoimmune diseases.
  • innate immune response cells such as macrophages, dendritic cells, Langerhans cells, and microglia sense the presence of a respective ligand ( stimulus-specific)
  • RNAs expressed from DNA by microarrays It has been clarified by kinetic analyses of RNAs expressed from DNA by microarrays that various combinations of the innate immunity ligands, combinations of antigens and innate immunity ligands, and the like lead to variety of changes to the processes and differences in actions thereof. This is because technological advances in kinetic analyses of DNAs and RNAs have permitted capturing changes, in time sequence thereof, of the expression process of innate immunity and lymphocyte immune system (the so-called acquired immunity), the apoptosis pathway, inflammation controls (anti-bacterial, anti-cancer) pathway, switching to an antiviral pathway, inflammation and inflammation control processes, and the like.
  • acquired immunity the so-called acquired immunity
  • inflammation controls anti-bacterial, anti-cancer
  • LPS is a ligand that activates TLR4 and MD2
  • imidazoquinoline and its derivatives are ligands that activate TLR7
  • flagellin is a ligand that activates TLR5.
  • imidazoquinoline and R-848 exhibit the same TLR activity, they produce different IFN- ⁇ types such that imidazoquinoline is used as a therapeutic agent for HPV (papillomavirus) while R-848 is used for HHV (herpes virus). This shows that even those agents with the same acceptor activity may exhibit different effects due to different ligands.
  • MDP muramyl dipeptide
  • MDP-Lys a derivative thereof, is a pharmaceutical agent as an adjuvant. This has recently received renewed attention for its aactivation of NLRs, but it is never used alone. It has been noted that a combination with other ligands such as LPS, Lipid A, or the like can elevate its ligand effect.
  • Patent Reference 1 has a finding that IL-10 is unexpectedly produced with a complex ligand made up of IL-12-producing bacteria and/or yeast with non-IL-12-producing bacteria and/or yeast.
  • complex ligands provide a synergistic effect absent in single ligands, in addition, permit the sites of action thereof to be diversified, thereby allowing the LPS (endotoxin) as a ligand to spread its risks, if it had any negative effect at all.
  • LPS endotoxin
  • a reason such as this has led to a strong need to develop a complexly-working ligand.
  • effects of such ligands have come to have an important significance after the discovery of flagellin's powerful adjuvant effects.
  • Modem health care is up against a wall.
  • Antibiotics that have been relied upon are ineffective for viruses and face serious problems of generating resistant- bacteria.
  • Hormones, such as steroids, and immunosuppressive drugs increase the risks of pathogenic and/or opportunistic bacterial infection through the weakening of the immunity, with their use failing to restore living organisms including the human life to normal.
  • treatment with inhibitors also suffers from adverse effects by hindering the normal living organism's systems.
  • a continued use of ineffective drugs is also escalating health care costs. Also from the standpoint of dismantling this wall, a safe and outstanding innate immunity-active ligand is being sought.
  • the present invention is aimed at providing a cytotoxicity-free and effective composition (ligand or adjuvant). Specifically, it is directed to providing a very safe composition (ligand or adjuvant) having a non-inflammatory disorder- inducing antibacterial effect, antiviral effect, anticancer effect, and an inflammation suppression effect including the runaway-suppression of CTL.
  • the present invention is aimed at providing a low molecular composition (low molecular ligand) that is readily permeable through the cell membranes.
  • low molecular composition low molecular ligand
  • the ease of cell membrane permeation means that it allows stimulating in a complex way the innate immune receptors present three-dimensionally in the endosomes on the cell surface and in the cell as well as the in the cytoplasm.
  • the present invention is aimed at providing a composition that stimulates three-dimensionally the innate immune receptor (ligand).
  • ligand innate immune receptor
  • the realization of a three-dimensional composition (ligand) permits one to expect a synergistic effect thereof, thereby further increasing safety.
  • by being low molecular it can also provide an immuno-activator that can also be readily absorbed through the intestine and skin. It is possible to realize a composition which cannot be attacked by antibodies because it is low molecular, making it also usable as an injectable agent.
  • the present invention is aimed at providing a composition that can be used as adjuvant.
  • Kaoru Mitarai one of the inventors of the present invention, collected aerobic soil fungi samples in a forest on a mountain near the city of Saiki City, Oita Prefecture, Japan, and cultured them over many years to obtain a strongly characteristic, stable group of symbiotic bacteria, which with more intensive research and experiments, resulted in the completion of the present invention.
  • the present inventors administered a low molecular degradation product obtained by degradation of bacterial cells from said symbiotic bacterial group to HIV-infected patients, when it was observed their conditions considerably improved, The CD4, an indicator of AIDS affliction, considerably increased in as short a period as a month, and all subjects gained vitality, which led us to name said group of symbiotic bacteria a "MRE symbiotic bacteria group".
  • MRE symbiotic bacteria group This means that the degradation products of the MRE symbiotic bacterial group enhance the innate immunity in patients, whereby the disease can be treated.
  • the present invention provides an innate immune enhancing composition capable of enhancing a subject's immunity, comprising the degradation product from the MRE symbiotic bacterial group.
  • an immune enhancing composition that enhances a subject's innate immunity by stimulating his innate immunity, comprising as an effective ingredient an immunostimulant generated by degrading at least one bacterium selected from an MRE symbiotic bacterial group consisting of Bacillus sp.(FERM BP-11209), Lysinibacillus fusiformis (FERM BP-11206), Bacillus sonorensis, Lysinibacillus sp. (FERM BP-11207), and Comamonas sp. ( FERM BP-11208).
  • said immunostimulant in such immune-enhancing composition is composed of at least 98 % by weight thereof or more of a hydrophilic low molecular substance of an average molecular weight of not more than 1,000 Da.
  • said immunostimulant in such immune-enhancing composition activates macrophages, natural killer cells, or natural killer T cells.
  • said immunostimulant in such immune-enhancing composition activates dendritic cells, microglial cells, Langerhans cells, or Kupffer cells.
  • said immunostimulant in such immune-enhancing composition activates epithelial cells, fibroblasts, keratinocytes, or osteoblasts.
  • said immunostimulant in such immune-enhancing composition causes the differentiation and activation of Th17 orTh1.
  • said immunostimulant in such immune-enhancing composition enhances IL-21 production.
  • said immunostimulant in such immune-enhancing composition is preferably obtained by incubating at least one bacterium selected from said MRE symbiotic bacterial group under culture conditions suitable for growth, placing the resultant culture medium under starvation conditions, and aerating.
  • said immune enhancing composition in such immune enhancing composition is used as an anti-inflammatory agent in a subject afflicted with allergic diseases or autoimmune diseases.
  • said allergic disease or autoimmune diseases can be selected from the group consisting of temporomandibular arthrosis, ulcerative colitis, atopic dermatitis, and allergic rhinitis.
  • the immune enhancement composition in such immune enhancing composition is used as adjuvant for vaccines against pathogenic bacteria or pathogenic viruses.
  • the pathogenic bacteria or viruses can be selected from the group consisting of opportunistic infectious bacteria, HIV, HCV, and HPV.
  • the immune-enhancing composition in such immune enhancing composition is used as a medicine or veterinary drug for treating the subject afflicted with a disease selected from the group consisting of liver cancer, prostate cancer, colon cancer, rectal cancer, lung cancer, pancreatic cancer, stomach cancer, malignant lymphoma, diabetes, hypertension, wound, ligament injury, bone fractures, low temperature burns, acne, floaters, bedsores, and urticaria, or for preventing said disease.
  • a disease selected from the group consisting of liver cancer, prostate cancer, colon cancer, rectal cancer, lung cancer, pancreatic cancer, stomach cancer, malignant lymphoma, diabetes, hypertension, wound, ligament injury, bone fractures, low temperature burns, acne, floaters, bedsores, and urticaria, or for preventing said disease.
  • the immune-enhancing composition in such immune enhancing composition is used as food or feed.
  • a method that produces an immune-enhancing composition that enhances a subject's innate immunity by stimulating innate immunity, said method comprising making available at least one bacterium selected from the MRE symbiotic bacterial group consisting of Bacillus sp.(FERM BP-11209), Lysinibacillus fusiformis (FERM BP-11206), Bacillus sonorensis, Lysinibacillus sp. (FERM BP-11207), and Comamonas sp. ( FERM BP-11208) and degrading said at least one of said bacteria made available.
  • MRE symbiotic bacterial group consisting of Bacillus sp.(FERM BP-11209), Lysinibacillus fusiformis (FERM BP-11206), Bacillus sonorensis, Lysinibacillus sp. (FERM BP-11207), and Comamonas sp. ( FERM BP-11208) and degrading said at
  • said step of degrading in such method is performed by incubating said at least said one bacterium made available under culture conditions suitable for growth, placing the resultant culture medium under starvation conditions, and aerating.
  • a method for treating a mammal afflicted with an innate immunity relateddisease, or for preventing said disease comprising making available a therapeutically effective amount of an immune-enhancing composition having as an effective component an immunostimulant produced by degrading at least one bacterium selected from the MRE symbiotic bacterial group consisting of Bacillus sp. (FERM BP-11209), Lysinibacillus fusiformis (FERM BP-11206), Bacillus sonorensis, Lysinibacillus sp. (FERM BP-11207), and Comamonas sp. ( FERM BP-11208) and administering a therapeutically effective amount of said immune-enhancing composition made available to the mammal.
  • an immune-enhancing composition having as an effective component an immunostimulant produced by degrading at least one bacterium selected from the MRE symbiotic bacterial group consisting of Bacillus sp. (FERM BP-11209), Lysinibacill
  • said mammal is a human.
  • said administering step is performed orally.
  • said administering step is performed parenterally.
  • parenteral administration step is preferably selected from intravascular administration, injection around or into the tissue, subcutaneous injection, intraocular administration, nasal administration, transdermal administration, and mucosal administration.
  • said innate immunity-relateddisease can be selected from the group consisting of allergic diseases or autoimmune diseases including temporomandibular arthritis, ulcerative colitis, atopic dermatitis, and allergic rhinitis; pathogenic bacterial- or virus-related diseases including those of opportunistic infectious bacteria HIV, HCV, and HPV; liver cancer, prostate cancer, colon cancer, rectal cancer, lung cancer, pancreatic cancer, stomach cancer, malignant lymphoma, diabetes, hypertension, wound, ligament injury, bone fractures, low temperature burns" acne, floaters, bedsores and urticaria.
  • allergic diseases or autoimmune diseases including temporomandibular arthritis, ulcerative colitis, atopic dermatitis, and allergic rhinitis
  • pathogenic bacterial- or virus-related diseases including those of opportunistic infectious bacteria HIV, HCV, and HPV
  • an immune-enhancing composition can be provided that effectively enhances immunity functions including innate immunity activity by making use of a low molecular immunostimulant produced by degrading bacterial cells of the MRE symbiotic bacteria group.
  • said MRE symbiotic bacteria group consists of Bacillus sp.(FERM BP-11209), Lysinibacillus fusiformis (FERM BP-11206), Bacillus sonorensis, Lysinibacillus sp. (FERM BP-11207), and Comamonas sp. ( FERM BP-11208), all of which are aerobic bacteria.
  • low molecular is meant a molecule having a molecular weight such that it is permeable through the cell membrane and capable of intracellularly affecting.
  • a low molecular immunostimulant can be obtained that is cell membrane permeable and composed of bacterial cell degradation products containing low molecular peptides, carbohydrate chains, glycolipids low molecular nucleic acid degradation products, by degradation of bacterial cells of the MRE symbiotic bacteria group comprising aerobic gram-negative bacteria and aerobic gram-positive bacteria with a group of lysosomal enzymes or a group of lysosomal like enzymes.
  • This immunostimulant which is cell membrane permeable, reaches out not only to the TLR receptors present on the cell surface, but also to the internal TLR receptors present in the endosomes, phagosomes or the like and microbial sensing NLR receptors and viral sensing RLR receptors, present in the depth of the cells, thereby providing the innate immunity response cells with a profound three-dimensional ligand stimulus.
  • the immunostimulant obtained by degradation of bacterial cells of the symbiotic bacterial MRE group is composed approximately 99% thereof of a hydrophilic substance with a molecular weight not more than 3,000 and is made up of components such as oligopeptides including MDP-like substances, oligosaccharide chain, single-stranded RNA, thereby providing a low molecular cytotoxicity-free ligand component not available with common digestive enzymes such as so-called protease.
  • the ligand a product from a multiple number of mixed bacteria, is multifaceted, simultaneously subjecting the innate immunity receptors to a variety of pattern recognition stimuli. It is the latter aspect that distinguishes the low molecular immune enhancing composition prepared from the MRE symbiotic bacteria group.
  • the "immune-enhancing composition" derived from the above-mentioned MRE symbiotic composition may for convenience be referred to as " MRE complex ligand" both of which mean the same or is synonymous.
  • ligand refers to a substance that causes the activation of receptors by binding specifically to the immune receptors; complex ligand refers to a mixture or conjugate of multiple ligands. What acts as an adjuvant is also included in the "ligand.”
  • the MRE complex ligand is capable of stimulating intra- and extra-cellular innate immunity receptors cells into activating immunocytes in the innate immune system, such as macrophages, dendritic cells, Kupffer cells, microglia cells, Langerhans cells, natural killer cells, and natural killer T cells, and epithelial cells on the mucosa and/or skin, fibroblasts, Paneth cells present in the intestinal tract.
  • innate immunity receptors cells such as macrophages, dendritic cells, Kupffer cells, microglia cells, Langerhans cells, natural killer cells, and natural killer T cells, and epithelial cells on the mucosa and/or skin, fibroblasts, Paneth cells present in the intestinal tract.
  • said activation produces a Type I interferon, the release of which then causes the neighboring cells to simultaneously produce therefrom antibacterial and antiviral substances.
  • the activation of immunocytes in the innate immune system by the MRE complex ligands in accordance with the present invention causes substances such as interleukin to be secreted into the blood and/or lymph.
  • substances such as interleukin to be secreted into the blood and/or lymph.
  • This allows the T-cell lymphocytes to differentiate and activate into states called Th1 and TH17 that enhance antibacterial, antiviral, and anticancer potency.
  • the MRE complex ligand in accordance with the present invention will allow TH17 and the like to produce IL-21 that functions to suppress autoimmunity.
  • the present invention is capable of providing an immune enhancing composition with a novel low-molecular immunostimulant having antibacterial, antiviral, anticancer, and anti-inflammatory effects as well ensuing autoimmunity suppressive effects thereof, thereby solving the problems described above in accordance with the present invention.
  • the present invention is specifically explained, where the instant invention, as set forth above, pertains to a discovery of a group of novel microbial symbiotic bacteria called a MRE symbiotic bacterial group that produces a highly effective immunostimulant by degradation of bacterial cells thereof without high concentration culturing, thereby utilizing it to provide an immune enhancing composition.
  • the MRE symbiotic bacterial group is made up of five unique bacteria consisting of aerobic gram- positive bacteria and aerobic gram- negative bacteria.
  • These groups of bacteria are not just that of mixture of naturally present bacteria, but are comprised of symbiotic bacteria consisting of 5 unique bacteria, which are bacteria that were the culmination of our continued long term culturing of a variety of soil bacteria along with bacteria adhered to marine products through the initial bacteria's intense competing struggle for survival till uncovering each other's roles in the competition, which bacteria underwent a change in a stable manner through their variation or evolution.
  • the five bacterial group that makes up the MRE symbiotic bacterial group comprise, aerobic gram-positive bacteria, Bacillus sp.(Accession Number FERM BP-11209; Identification Number MK -005), Lysinibacillus fusiformis (Accession Number FERM BP-11206; Identification Number MK -001 ), Bacillus sonorensis, ( Accession Number FERM BP-11206; Identification Number MK -004) Lysinibacillus sp. ( Accession Number FERM BP-11207; Identification Number MK -002), and aerobic gram-negative bacteria Comamonas sp. ( FERM BP-11208; Identification Number MK -003).
  • Accession Number FERM BP -11206 (transferred from Accession Number FERM P- 21548 , deposited March 19, 2008), Accession Number FERM BP - 11207 (transferred from Accession Number FERM P- 21549 , deposited March 19, 2008); Accession Number FERM BP -11208 (transferred from Accession Number FERM P- 21550 , deposited March 19, 2008); and Accession Number FERM BP - 11209 (transferred from Accession Number FERM P- 21760, deposited February 2 2009 ) are the bacteria deposited at International Patent Organization Depository, Advanced Industrial Science & Technology, Japan, an independent corporation.
  • the MRE symbiotic bacteria group in accordance with the present invention can be grown under any conditions if under viable circumstances; specifically it is possible to grow under culture conditions commonly used in the field of molecular biology.
  • said bacteria can be incubated in a culture aeration tank, with aeration, in the presence of nutrients: 10kg fish meal, 10kg rice bran , 5kg oil meal, 1kg of broth along with minerals such as magnesium sulfate, and the like, at pH6.0 to 6.8, and 25°C to 35°C incubation temperature, with appropriate aeration.
  • said nutrient source is 5kg fish meal, 5kg rice bran, oil meal 2.5kg, and 0.5keg broth.
  • the culture conditions for the MRE symbiotic bacterial group in accordance with the present invention include, but not limited to, the terms of these settings.
  • the MRE symbiotic bacteria group in accordance with the present invention has progressed to the stage of active gene exchanging, undergone a mutation, and evolved into an enigmatic bacterial cell body close to bacterial fusion as with the Bacillus sp.
  • Respective bacteria that make up the MRE symbiotic bacteria group have the following 16Sr DNAs.
  • the first method is that of degrading the MRE symbiotic bacteria group with a group of lysosomal enzymes thereby obtaining a low molecular complex ligand.
  • the second method is that of obtaining a low molecular complex ligand by degrading MREE symbiotic bacteria group with primitive lysosomal homologous mother cell lytic enzymes that appear in the step of spore formation of the MRE symbiotic bacteria group
  • catabolic enzymes such as ordinary digestive enzymes and common proteases fail to provide ligands with molecular weights required to permeate the cell membrane.
  • lysosomal enzymes are enzymes involved in autophagy that breaks down aged organelles in cells and rejuvenates them and those that appear at the final stages where cancer cells and/or viral infected cells undergo apoptosis, which are particularly known to be active when degrading the bacteria that invaded into the cell in endosome cells.
  • Said homologous enzyme group is known to emerge in a processs of autophagy and/or apoptosis in animals, plants, and microbes. This is also a group of enzymes called processing enzymes in plants that emerge within the vacuoles or during fruit formation. These groups of enzymes would break down the bacterial cells down to low molecular substances small enough to be permeable through the cell membrane.
  • catabolic enzymes including nucleases such as ribonuclease, and deoxyribonuclease, and the like; proteolytic enzymes having powerful and versatile capabilities such as collagen-degrading enzyme cathepsin L, aspartic proteases cathepsin D and cathepsin E; cysteine proteases cathepsin K, cathepsin B, and cathepsin S; serine protease cathepsin G; aminopeptidase cathepsin H, and the like; arylsulfatase , ⁇ -glucurosidase,esterase, and acid phosphatase; carbohydrate chain degrading enzymes such as sphingolipid-degrading ⁇ -galactosidase, ⁇ hexosaminidases A and B, allyl sulfatase A
  • lysosomal enzymes are active when slightly acidic (pH6.3 to pH6.8) with their activity elevated at high temperature range (38°C to 42°C) where the activity of ordinary digestive enzymes is suppressed. Furthermore, among them are many having high degradation potency, some having the potency as high as 5,000 to 10,000 times that of the ordinary digestive enzymes.
  • the first method to obtain an MRE complex ligand from the MRE symbiotic bacterial group in accordance with the present invention is one which uses a combination of a cell wall-lytic enzyme and cathepsins from among these lysosomal enzymes with a nuclease.
  • the cell wall-lytic enzymes use is made of muramidase, mucopeptide hydrolase , and the like; for the cathepsins, cathepsin B, cathepsin D , cathepsin L, cathepsin K, or papain; for the nucleases, ribonuclease, and deoxyribonuclease. While these enzymes can be created by gene recombination into a plasmid or DNA, they can also be obtained by having fishes (including eel) autolyzed under sterile conditions at elevated temperatures (38°C to 45°C), in a moist environment. They can also be obtained by using a group of lysosomal enzymes produced when fruits such as papaya ripen.
  • the first method is advantageous in that the bacterial cell degradation can be adjusted by changing the enzyme blend ratio
  • a second method to obtain a low-molecular complex ligand from the MRE symbiotic bacteria group is one which uses, as is, a primitive mother cell lytic enzyme released during the spore formation step. Although the enzyme blend cannot be varied, excellent ligands can be mass produced at a very low cost.
  • This method is one for obtaining a low-molecular complex ligand using the mother cell lytic enzymes released in connection with spore formation of the MRE symbiotic bacteria group.
  • mother cell lytic enzyme group or “mother cell lytic enzyme” as used in the present invention refers to the lysosomal homologous enzymes produced during a spore formation step of bacterial cells.
  • Preparation of a low molecular MRE complex ligand in accordance with the present invention calls for, at its first stage, charging a blend culture medium with a MRE symbiotic bacterial group and incubating under the culture conditions involving the addition of, as bacterial nutrients, fish meal, rice bran, oil cake, broth and the like, minerals such as magnesium sulfate, at a culture pH 6.0 to 6.8 and a culture temperature at 25°C to 35°C , along with sufficient aeration (aeration: 0.1mg / L to 1.0mg / L dissolved oxygen concentration).
  • the system After the growth and stabilization of the MRE symbiotic bacterial group is ttained, the system is placed under a starvation condition with the nutrients cut off, followed by continued aeration, whereupon the depletion of nitrogen components triggers a sporulation (internal sporulation).
  • a stable quality product can be obtained if the vegetative cells prior to sporulation are transferred to another aeration tank. In this manner it is made possible to degrade the bacterial cells of the MRE symbiotic bacteria group down to low molecular products.
  • the bacterial cell degradation process in accordance with the present invention is further detailed, wherein blended bacterial cells which have built mutual bacterial symbiotic relationships and reached a stable symbiotic body's vegetative cell state, along with the culture containing digestive enzymes secreted from the vegetative cells thereof, are separated into another aerating culture vessel. Aeration is continued in the aerating culture tank with no nutrients other than silica. Autophagosomal homologous mother cell lysis of the MRE bacterial group in a symbiotic state begins, and the bacterial vegetative cells undergo bulk degradation and vanish while releasing lysosomal homologous mother cell lytic enzymes.
  • the MRE symbiotic bacteria group permits a sufficiently effective MRE complex ligand to be prepared even without high concentration production, but it is also possible to carry out high concentration production.
  • repetitious formation of vegetative cells and sporulation results in the vegetative cells absorbing the complex ligand, adversely affecting the yield and in an increase in the production of antibiotics, such that it is important to perform the sporulation all at once after a high density culturing.
  • providing a silica nutrient further improves efficiency.
  • the product obtained by sporulation all at once was essentially free of any antibiotics, below a detectable level.
  • the low molecular complex ligands prepared in this manner contain oligopeptides, single-strand RNA degradation products, oligosaccharides, glycolipids and MDP (muramyl dipeptide) -like substances, and flagellin degradation products, having a molecular weight distribution as shown below.
  • Table 2 shows the molecular weight distribution of a MRE complex ligand.
  • Table 2 Molecular weight distribution of MRE complex ligand Range of molecular weight Peak area (%) Not lower than 10,000 Trace 3,000 to 10,000 Trace 1,000 to 3,000 1.5% 500 to 1,000 1% Not higher than 500 97.5% Total 100%
  • the MRE complex ligand of this invention is composed of hydrophilic low molecular substances, 1,000 or less, including oligopeptides, oligosaccharide chains, oligo level nucleic acids, oligo level glycopeptides and glycolipids.
  • the resultant low molecular complex ligand obtained in this way has no toxicity, such as an endotoxin, and being low molecular makes it free from a direct antibody attack.
  • the MREcomplex ligand is such that even with an endotoxin test capable of detecting the presence of LPS and peptidoglycan, their presence was below the detection limit, demonstrating the absence of the LPS and peptodiglycan.
  • an endotoxin test capable of detecting the presence of LPS and peptidoglycan, their presence was below the detection limit, demonstrating the absence of the LPS and peptodiglycan.
  • a cell dysfunction test used for performing an activity test for macrophages, natural killer cells, and the like showed that there was no cell dysfunction up to the limiting concentration at which a breakdown occurs due to osmotic pressure.
  • said ligand also has cleared an acute toxicity test using a rabbit.
  • a group of immunocytes in the human blood was separated; and the production of each cytokine by the MRE complex ligand was examined using an antibody.
  • the stock solution containing the MRE complex ligand as used contains 80 ⁇ g / ml of the MRE complex ligand-containing low molecular components.
  • the amount of TNF- ⁇ production was determined using real-time PCR, resulting in the production 90.46 times the normal level as shown in Table 3.
  • Med stands for no stimulus.
  • LPSp is for an endotoxin stimulus; MRE 1/ 10 is for a 10- fold dilution of the MRE complex ligand-containing stock solution; and the same thereafter for 100- and 1000-fold dilutions.
  • the LPSp concentration for comparison is 100 ng / ml.
  • the MRE complex ligand activates the immunizing process involving innate immunity activation and the ensuing antiviral, antibacterial, anti-inflammatory effects, and tissue repair thereby restoring the living body back to normal. It could be something called a breakthrough ligand or adjuvant.
  • TLR innate immune receptors
  • the MRE complex ligand activates a group of NLR innate immune receptors including TLR-2, TLR-7, TLR-8, and NLR-2, which are innate immunity receptors.
  • TLR-2 which is an innate immunity receptor present on the cell surface, senses Gram-positive bacterial peptidoglycan, lipoteichoic acid, lipoprotein, viral glycoprotein, fungal polysaccharides, and the like.
  • Active in the MRE complex ligand is an MDP-like peptidoglycan degradation product having a molecular weight of not more than 1000.
  • TLR-7 and TLR-12 are receptors for sensing single-stranded RNA and also for sensing a low molecular substance from bacterial cell degradation of bacteria and/or viruses.
  • the MRE low-molecular complex ligand of the present invention is thought to sense single-stranded RNAs having a molecular weight of not more than 1000. This fact means that the present invention can be expected to have a high adjuvant effect against RNA viruses.
  • NLRs including NLR-2 which are primitive receptors present intracellularly, are able to detect low-molecular bacterial or viral degradation products.
  • NLR-2 is a receptor that expresses specifically to APCs (antigen presenting cells) such as macrophages, dendritic cells, Langerhans cells, and Kupffer cells.
  • APCs antigen presenting cells
  • the group of NLR receptors activated by the MRE low molecular complex ligands can sense MPD-like substances and the like low molecular ligands. This again permits one to expect the ligand to operate very effectively as an adjuvant.
  • the MRE low molecular complex ligand of the invention are sensed three dimensionally by three innate immunity sensors, which are receptors present on the cell surface, receptors present in the endosomes, and receptors present intracellularly.
  • the NO production capability with an unstimulated culture medium is 0.446 ⁇ M
  • the production capability with a 10 fold diluted solution of the MRE complex ligand -containing stock solution is 24.059 ⁇ M, a value greater than that of 18.712 ⁇ M obtained with a 0.1ng/ml LPSp solution.
  • the viable cell count falls with an increasing concentration of LPS, as shown in Table 5, whereas with the MRE complex ligand, the viable cell count increases as its concentration increases.
  • This macrophage cell viability test entailed determining the macrophage plate adhesion ratio as an index for the viable cell count by staining with crystal violet and measuring the absorbance at 570 nm.
  • the process to apoptosis is the process that gives rise to a cascade of apoptosis, in which FADD triggers apoptosis, leading to activation of caspase 8 enzyme and/or caspase 10 enzyme, and induction of apoptotic degradation enzymes such as cathepsin D and cathepsin B, which are execution unit enzymes.
  • apoptosis-inducing genes AP1s is at a normal level; apoptotic suppression gene JUN is expressed (6.77409 times the normal level); and also expressed is gene SOD2 (8.99963 times the normal level) that guards against the release of apoptosis-inducing cytochrome C from the mitochondria, thereby blocking apoptosis.
  • the antiviral substance production process is one of the processes of original innate immunity, in which a cascade of process activation steps propagates: stimulation of TLR 3, TLR7, TLR8, and RGR leading to the expression of TRAM and TRIF, followed by IKK activation, and activation of IRAK 3, IRAK 7 and the like, resulting in the production of Type I IFN- ⁇ (now 13 types are known ) and IFN- ⁇ to be released extracelluarly.
  • This process is believed to be a mechanism built in the unicellular age; in humans, the release of these interferons causes a concurrent release of ISG antiviral substances (several hundred types are known) from neighboring cells such as epithelial cells and mucosal cells.
  • ISGs which are antiviral substances produced dependent on the stimulation pattern.
  • influenza leads to the release of antiviral substances such as IFIT1, GIP3, GIP2, OAS1, M1X1, IFIH1, IFIT3, RIG-I, GBP1, LAMP3, IRF7, ISGF3G, WARS, PSMBS6, BTC, SOCS1, and SERPING 1 thereby exhibiting antiviral effects in response to viral mutation.
  • the MRE complex ligand by having been activated prior to this process, makes it possible to produce type I interferon immediately and powerfully when the host is infected with viruses such as influenza, thereby effectively reinforcing antiviral potency. In other words its adjuvant effect is at a very high level.
  • Table 6 reveals how much type I interferon gene expression increases with the MRE complex ligand, in the absence of viral infection, using a comprehensive DNA kinetic analysis.
  • Type I Interferon production (M ⁇ ) IL28a 1.84637
  • IFN- ⁇ ILNA6 1.67936
  • IFN- ⁇ IFN5 1.27655
  • IFN- ⁇ IFNB1 1.41305 IFN- ⁇
  • MRE complex ligand is seen to give significant increases in IFN- ⁇ and IFN- ⁇ .
  • the IFN- ⁇ production also has further increased. (See Figure 3 ) As just described herein, the MRE complex ligand is believed to have no side effects as an antiviral adjuvant.
  • macrophage-sibling dendritic cells microglia cells, Kupffer cells, Langerhans cells, fibrosis cells as well as epithelial cells and keratinocytes that express the same innate immunity receptors, and in addition, innate immunity response cells located in each of the organs including the trachea, the gastrointestinal tract, and the urinary tract.
  • This process is the process from the innate immunity receptor stimulation to production of interleukins that stimulate T cells, swhich is made up of the following two-stage process:
  • NLR receptors when stimulated with a ligand, the NLR receptor pairs and RICK binds thereto through mediation of NLR's CARD, leading to ubiquitination of the RICK, followed by further binding of complexes such as TAK1, MEMO, and the like, thereby activating IKK ⁇ , separating I- ⁇ B from NF- ⁇ B, and activating NF- ⁇ B.
  • complexes such as TAK1, MEMO, and the like
  • chemokines including IL-1 ⁇ , TNF- ⁇ , and IL-8.
  • I ⁇ B- ⁇ joins NF- ⁇ B and AP-1 which operate on DNA, thereby producing IL-12p40, IL-6, and others, resulting in control of the T lymphocyte immunity.
  • Table 8 shows the results from a comprehensive DNA kinetic analysis of the first stage process of gene expression when stimulated with the MRE complex ligand using human macrophages. This result is also consistent with that from a real-time PCR analysis.
  • JUN and/or SOD2 are expressed to block TNA- ⁇ and the like from inducing apoptosis of the host's own cells, so as to prevent, with SOCS3, the inflammation from progressing excessively in response to more-than- necessary stimuli from the innate immunity receptors.
  • PDLIM capable of ubiquitinating and degrading excess NF- ⁇ B, did not need to operate so that it was at a normal level of 1.0721337.
  • NFKBIA 8.42561 I- ⁇ B production NFKBID 2.10104 I- ⁇ B production IKBKE 1.4477 I- ⁇ B production NLRP4 1.35103 Suppressing NF- ⁇ B JUN 6.77408 Apoptosis suppression SOD2 8.99963 Apoptosis inhibition SOCS3 2.62260 Inflammation control PDLIM2 1.07213 Degrading NF- ⁇ B
  • TNFAIP6 is a gene that strongly suppresses NF- ⁇ B, has a strong anti-inflammatory action that controls the TNF- ⁇ production, and expresses at a high value of 37.0424 times the normal level.
  • TNFAIP3 is also a gene that suppresses the expressions of NF- ⁇ B and AP-1 and has also a high value of 18.8308.
  • JUN, SOD, SOCS3 are operating is as was already described.
  • the second stage process is triggered by I- ⁇ B- ⁇ (trigger). Since a large number of NF- ⁇ B and AP-1 are already in operation as bound to nucleic acids, delayed inflammation suppression genes are made more active thereby diminishing the fresh supplies of NF- ⁇ B and AP-1.
  • the newly-created IkB- ⁇ triggers and binds to the NF- ⁇ B which is already bound to nucleic acids and together with AP-1 which is similarly already bound to nucleic acids, produce and secrete four groups of cytokines that control T cells.
  • the M1 activation produces cytokines "IL-12p40,” “IL-6 and IL-23p19”; M2 activation, "IL-4,””TGF- ⁇ , "IL-2” and the like. This is followed by binding to p35, which is constantly being produced intracellularly, thereby turning IL-12p40 into TL-12 and IL-23p19 into IL-23.
  • Table 10 shows how the gene expression takes place, starting the second stage by stimuli with the MRE complex ligand.
  • the I- ⁇ B- ⁇ trigger rapidly increased to 20.015 times the normal level and the NF- ⁇ B and AP-1 had their production fall by virtue of the enhanced expression of inflammation suppression genes.
  • the MRE complex ligand brings macrophages and siblings thereof such as microglia cells, dendritic cells, Langerhans cells, Kupffer cells, and the like to M1 activation, thereby exerting antibacterial and antiviral effects. It also invigorates phagocytic activity on apoptotic cancer cell debris.
  • M2 activity is the posttreatment process for suppressing inflammation and activating fibroblasts so as to repair the tissues destroyed by inflammation and the like. In light of the Examples it is suggested that also in M2 does the MRE complex ligand play important roles.
  • the MRE complex ligand has excellent properties in that apart from the conventional M1 activity processes of expressing antibacterial, antiviral or anticancer actions, it permits concurrently running an anti-inflammatory process with inflammatory diseases including allergic diseases and autoimmune diseases.
  • Macrophage M1 activity effects the induction of Th17 and Th1, while macrophage M2 activity effects the induction of Treg or Th2. Further, it has been found that the MRE complex ligand in accordance with the present invention is able to induce Th17 and Treg and to suppress the induction of Th 1 and Th2.
  • IL-12p40 and IL-23p19 among a group of cytokines produced through a series of processes due to the stimulation of innate immune receptors, bind to intracellularly-residing p35 thereby turning into IL-12 and IL-23, respectively.
  • Th 1 and Th17 processes following the M1 activity provide antibacterial, antiviral, and anticancer and allergy suppressive actions; Th 2 following M2 activity leads to allergic reaction processes; and the Treg process provides inflammation suppression, tissue repair, and autoimmune suppression (immune tolerance) actions.
  • Th1 produces IFN ⁇ by the action of IL-12.
  • Production of free radicals such as NO thereby enhances intracellular antibacterial potency.
  • a process to eliminate intracellular parasitic bacteria and/or viruses is induced by an enhancement of the activity of CTL (killer T cell), CD8-T cells and an increase in cell-mediated immunity that kills cancer cells and virus-containing cells. This is primarily a non-antibody-dependent antibacterial and anti-viral process. It is also a process causing cytotoxic autoimmune cell diseases and/or delayed-type allergic diseases.
  • Th2 produces IL-4 and IL-13 and causes B cells to release IgE antibodies through mediation of CD4-T cells.
  • IgE acts to eliminate large organisms such as parasites by secreting histamine from mast cells and/or basophils.
  • it is also known to produce IL-5 thereby releasing EPO from eosinophils and induce delayed allergic conditions such as stuffy noses and the like.
  • Th17 has IL-1 and/or IL-23 operate to produce IL-17A, IL-17F, IL-22, IL-21, and the like.
  • humoral immunity extracellular bacteria and fungi are eliminated by a vigorous production of antibodies.
  • neutrophilic phagocytosis causes antimicrobial substances such as defensin to be released primarily from epithelial cells and neutrophils, and strengthens the epithelial barrier through an extracellular matrix remodeling.
  • Treg which characterizes intestinal immunity, is induced by TGF- ⁇ , producing IL-10, which is believed to be a process of inducing inflammation suppression and/or immune tolerance.
  • TGF- ⁇ producing IL-10
  • IL-10 which is believed to be a process of inducing inflammation suppression and/or immune tolerance.
  • the intestinal tract with abundant enterobacteria is normally held in the Treg state so as to prevent lymphocyte immunity from running away, where its defense strength is delegated to the innate immunity of the epithelial cell lining and /or Paneth cells, and at the same time the enterobacteria are controlled by secretion of IgA, a secreting antibody, into the intestine through Peyer's patches.
  • Treg is also known to suppresses inflammation by bringing into apoptosis, or suppressing, B cells that secrete allergy-causing Th1 and /or IgE and CTL(killer cells) that cause autoimmune diseases.
  • MRE complex ligand which follows the macrophage M1 activity process, as shown in the results of the previous Table, IL-23p19 and IL-12p40 increase;IL-4 does not change and TGF- ⁇ v rather tends to fall off, showing that the ligand activates Th17 and Th1 but not Th2 and Treg.
  • the MRE complex ligand suppresses inflammation as in ulcerative colitis and Crohn's disease such that the process of the Trig being restored to normal with the MRE complex ligand in the actual intestinal tract is also believed to contribute to inflammation suppression and /or immune tolerance.
  • the MRE complex ligand-containing solution increased IL-23 production to 1.92 times the normal level even on average; it was interesting to note that seemingly the older the subject the greater the IL-23 value. Accordingly, the MRE complex ligand increases the Mlactivity of macrophages or the like , thereby activating Th17 and Th 1 and elevating cell-mediated immunity and humoral immunity so as to exert antibacterial, antiviral, and anticancer effects. This is consistent with clinical trials, including those shown in the Examples.
  • MRE complex ligand very strongly activates the genes of inflammatory cytokines such as IL-1 ⁇ ,TNF- ⁇ , IL-8 and the like and activates Th-17 and Th1 processes, nevertheless for some reason, a beverage containing the MRE complex ligand significantly improves inflammation in cases of jaw inflammation, ulcerative colitis, atopic dermatitis, and the like.
  • the inventors made intensive studies focused on the comprehensive DNA kinetic analysis of macrophages using a DNA array and the time sequential progress of cytokine production using human blood.
  • the MRE complex ligand expresses a novel immune process, not existing with the conventional ligands such as LPS.
  • the first point is a suppressive action on IL-18 production by the MRE complex ligand.
  • the current DNA kinetic analysis revealed, as shown in Table 12, that the MRE complex ligand suppresses the IL-18 production. This was a remarkable discovery.
  • IL-18 is a cytokine closely related to inflammatory diseases, which generates caspase 1 enzyme when stimulated by a conventional M1 ligand such as LPS; the caspase 1 cleaves an IL-18 precursor, producing IL-18.
  • IL-18 then acts, in the presence of IL-12, on Th1 cells and /or NK cells, thereby strongly inducing IFN- ⁇ production. This is because IL-12 increases IL-18 receptors in the Th1 cells and NK cells.
  • IFN- ⁇ whichTh1 cells produce, promotes the production of IL-12 and /or 11-18 from macrophages, in turn stimulating Th1 cells whereby a cycle will be formed to sustain and increase inflammation.
  • both IL-12 and IL- 18 will rise.
  • IL-18 known characteristics of IL-18 are that upon a simultaneous excessive production of IL-12 and/or IL-23 and IL-18, severe organ damage and autoimmune diseases develop in the intestines and the liver.
  • the production of IL-4 and /or IL-13 is promoted where activation of a Th2 process leads to IGE production and histamine is released from the mast cells and /or basophils.
  • EPO is released from eosinophils causing inflammatory conditions such as nasal congestion.
  • IL-18 by itself directly stimulates, with no IGE mediation, mast cells and / basophils thereby releasing histamine or inducing IL-4 and IL-13 production.
  • IL-13 is also known as a cytokine that induces bronchial asthma and/or pulmonary fibrosis (See Non-Patent Reference 4).
  • Th1 cells when Th1 cells are acted upon by IL-18 and TL-12 that originally activates Treg, the Th1 cells are transformed into cells called the super Th1, releasing IL-13, which is originally a Th2 cytokine, thereby causing bronchial asthma and lung fibrosis. It is believed that in the case of atopic dermatitis caused by Staphylococcus aureus, an aggravation also occurs by a similar mechanism.(Non-Patent Reference 6).
  • IL-18 as a trigger is considered to switch Th1 cells, Th2 cells, Th17 cells, and the like between two states: inflammation and non-inflammation.
  • Th1 cells cause autoimmune diseases and Th2 cells cause allergic diseases.
  • Th2 cells cause allergic diseases.
  • cell mediated immunities such as intracellular antibacterial activity, antiviral activity, and anticancer activity are reinforced.
  • I1-4 and /or IL-13 and the like are produced in the co-presence of IL-18 thereby inducing allergic diseases.
  • a secretory immune process activates so as to secrete IgA from the intestinal tract and/or mammary gland.
  • an IL-18 trigger results in a direct stimulation, with no IGE mediation, of the mast cells and / basophils thereby causing allergic diseases. It may be thought that also with Th 17, an IL-18 trigger causes a diversion into a process of promoting inflammation and that of suppressing inflammation.
  • IL-17A and/or IL-22 activating a humoral immune process. It is also known that an excessive production of IL-17 is highly inflammatory, developing autoimmune diseases such as chronic arthritis, multiple sclerosis, ulcerative colitis, Crohn's disease, and psoriasis. IL-22, closely associated with psoriasis, causes excessive production of HNP-3, an ⁇ -defensin.
  • IL-17 activates a humoral immune process for elevating antibacterial and antiviral potency, such as vigorous antibody production and defensin secretion.
  • IL-17 is known to promote the production of ⁇ -defensins, such as HNP 1 to 2 and HNP4 to 6 from neutrophils as well as the production of ⁇ -defensins, hBD1 to 4, from epithelial cells (See Non-Patent Reference 7 )
  • the second point of the notable action of the MRE complex ligand is its action to increase the IL-21.
  • the MRE complex ligand by virtue of suppressing the production of inflammation-causing IL-18, is expected to effect the suppression of inflammations including allergies and /or autoimmune diseases, which actually is consistent with the clinical trial results for inflammatory diseases.
  • IL-21 is a cytokine that elevates the anticancer activity by multiplying the secretion of antiviral agents and/or natural killer cells (See Patent References 4 and 5; Examples 8 to 13); separately, it also reportedly functions to cause apoptosis of T cells and B cells that cause autoimmune diseases (See Patent Reference 3).
  • IL-21 It is known for IL-21 that there is a process wherein IL-12 is released from the macrophage M1 cells; and the IL-12 acts directly on NKT cells (natural and killer T cells), thereby inducing IL-21 production (See Patent Reference 2).
  • Th17 activation induces the production of IL-21 from Th 17(See Non-patent Reference 3) and furthermore that IL-21 has a cycle that promotes differentiation of Th17 cells and that secretes IL-21 on stimulation of IL-1 and /or IL-2 (Non-Patent Reference 8).
  • the MRE complex ligand may be said to primarily produce IL-21 due to a Th17 non-inflammatory activation, and also secondarily to make use of an IL-121 production route due to an NKT cell activation via IL-12.
  • vertebrates, including humans are constantly subjected to stimuli from ligands other than the MRE complex ligand such that these two IL-2 production routes would also constantly vary. In such cases the MRE complex ligand assumes characters more strongly as an adjuvant and will exhibit its excellent adjuvant effects.
  • the MRE complex ligand had 1.2 times the normal expression of cathepsin E genes that appear when the macrophages suppress inflammation.
  • the MRE complex ligand provides the amazing actions of calming inflammation while maintaining its activities toward the antibacterial, antiviral, and anticancer immunity processes.
  • the MRE complex ligand expresses FGF2, one of the cytokines that promote tissue repair, at 2.525 times the normal level and is capable of rapidly restoring tissue damage, which is also consistent with clinical trials.
  • the MRE complex ligand of the invention performs a three-dimensional M1 activation of intra- and extra-cellular innate immune receptors including macrophages, microglia, dendritic cells, Langerhans cells, Kupffer cells, epithelial cells, keratinocytes, fibrotic cells, thereby causing the secretion of antibacterial and antiviral substances such as defensins and ISG as well as activating NK cells and NKT cells to cause apoptosis of cancer cells and virus-containing cells.
  • NF- kB activates NF- kB in two stages and secretes chemokines including IL-1 ⁇ , TNF- ⁇ , and IL-8, producing at the same time multiple inflammation suppressors and /or mitochondrial SOD, thereby inhibiting cell damage due to TNF- ⁇ and the like. Further it causes the production of T lymphocyte cytokines, such as IL-6, IL-12, IL-12 and IL-23, differentiation of naive T cells into Th17 cells and Th1 cells, and activation of them. In addition the MRE complex ligand suppresses IL-18, a trigger of various diseases, and switches Th17 activity and Th1 activity for induction into non- inflammatory processes. This switching causes Th17 to produce IL-21 and IL-17.
  • chemokines including IL-1 ⁇ , TNF- ⁇ , and IL-8, producing at the same time multiple inflammation suppressors and /or mitochondrial SOD, thereby inhibiting cell damage due to TNF- ⁇ and the like. Further it causes the production of T lymph
  • a cycle operates where the IL-21 differentiates naive T cells into non- inflammatory IL-17.
  • IL-17F enhances antibody production with activation of a humoral immune process free from inflammation, and in addition, secretes directly the innate immunity antibacterial substance defensins and/or antiviral substances, thereby exerting antibacterial and antiviral potency effects.
  • the release of IL-12 causes IL-21 production from NKT cells without IFN- ⁇ production.
  • IL-21 in these cases exerts anticancer activity by directly increasing NK cell activities and in addition, causes apoptosis of autoimmune disease-causing T cells and /or B cells so as to suppress autoimmunity.
  • macrophage activation invigorates phagocytosis thereof to facilitate the removal of wastes and foreign material in the blood, lymph, and tissue body fluid.
  • the vaccines used in combination with the MRE complex ligand as an adjuvant include antiviral and anti-bacterial vaccines such as vaccines for influenza, plague bacillus, and the like; new generation LPS, peptidoglycan, lipoarabinomannan, zymosan, lipopeptides, lipoteichoic acid, RSV-F protein, fibronectin EDA domain, HSP60, flagellin, unmethylated CpG DNA, double-stranded RNAs, polyinosinic polycytidylic acid, imidazoquinoline compounds, ⁇ -glucan, Maruyama Vaccine, mycobacterium bovis; and TLR ligands in OK-432 and the like, as well as vaccines containing bacterial cell components containing viruses bound thereto.
  • antiviral and anti-bacterial vaccines such as vaccines for influenza, plague bacillus, and the like
  • new generation LPS new generation LPS, peptidoglycan, lipoarabinomannan, zymos
  • the MRE complex ligand can be used in blends with other immune ligands and/or immune adjuvants.
  • it is highly compatible with carbohydrate chain type immune active ingredients in that degrading substances containing carbohydrate chain components such as lingzhi (reishi) mushroom, cordyceps, chaga mushroom (Inonotus obliquus), chitin chitosan, Agaricus and the like, to a level not more than a molecular weight of 8,000, followed by adding to the MRE complex ligand, exhibited a 49% to 60 % increase in macrophage activity (viable cell count and NO production ).
  • This shows that the MRE complex ligand can function as an excellent adjuvant.
  • Cancer therapies with the combined use of a substance that restores apoptosis to cancer [cells] such as koto-sugi (family Taxaceae), resveratrol, quercetin or the like is also very effective.
  • DNA kinetic analysis using a DNA microarray was carried out using the Human Gene 1.0 ST Array, Bio-Matrix Research.
  • RNAs obtained by this procedure were used to analyze the comprehensive gene expression.
  • the MRE complex ligand of the invention activate innate immunity and the subsequent lymphocytic acquired immune processes, but also it serves to suppress inflammation and enhance tissue repair potency. Accordingly, the MRE complex ligand of the invention has antibacterial, anticancer, antiviral, anti-inflammatory, tissue repair, and waste removal capabilities.
  • the MRE complex ligand of the invention has a stimulation potency 7.54-fold that of LPS (endotoxin), which has powerful innate immune activation potency. It was also established that at the same time, numerous anti-inflammatory components appear and the mitochondrial SOD is produced such that TNF- ⁇ does not increase in the blood and no cell damage is caused. It was also confirmed that the MRE complex ligand has excellent specific properties in that it has no cytotoxicity in contrast to LPS. A fact was shown that on the one hand, with LPS, as its concentration increases, the viable cell counts of macrophages fall; on the other hand, as the MRE complex ligand concentration increases, viable cell counts of macrophages increase.
  • the MRE complex ligand of the invention which is a low polarity, essentially electrical charge-free, oligo level low molecular ligand, is readily permeable through the cell wall so that it can stimulate and activate three-dimensionally and complexly not only the TLR receptors expressed on the cell surface but also NLR and/or RLR receptors expressed in intracellular endosomes and/or present in the cytoplasm.
  • Such low molecular ligand can be readily absorbed through the intestinal wall, and can also be used, after purification thereof, for intravenous injections because they are not attacked by antibodies. This property provides synergistic effects with specific properties not seen with other ligands, along with the property of protecting cells and increasing cell viability with an increasing concentration thereof.
  • Tables 13 and 14 show data where human macrophages were subjected to a diluted solution of the MRE complex ligand-containing stock solution prepared in "Example 1";and the NO production and the viable cell count for the macrophage were determined.
  • the results of determination show that the NO production with the MRE complex ligand-contaminating stock solution was greater than that with LPS; specifically the NO production with the 300-fold dilution thereof was as high as 2.19 fold.
  • the MRE complex ligand-containing stock solution gives an increase in NO
  • the macrophage viable cell count index increases with the MRE complex ligand-containing stock solution, which shows an opposite trend with the LPS in which the index fall off.
  • the MRE complex ligand, causing no cell damage is rather said to be an immune-activator with a macrophage activation effect, raising the viability of the macrophage.
  • the MRE complex ligand-containing stock solution is not directly bactericidal was established by a growth inhibition check test using Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus, Candida albicans, and Black aspergillus. Therefore, the MRE complex ligand-containing stock solution contains no bactericides including antibiotics; and all the antibacterial effects seen in clinical trials can be ascribed to its immunocompetency.
  • the antibacterial effects of the MRE complex ligand in accordance with the present invention can be summarized as follows: That is, the first point is its antibacterial effect due to an increase in macrophage phagocytosis; the second point is an antibacterial effect due to a release of antibacterial substances including defensins from macrophages, epithelial cells, keratinocytes, and the like. Specifically, it was confirmed that the release of PTX3, an antibacterial substance, from macrophages is 7.27-fold that of the normal level.
  • the third point is an antibacterial effect brought out by an increase in the phagocytic activity of neutrophils through Th17 activity, where phagocytosed bacteria are killed by HNP1 to 6, ⁇ -defensins present in the azurophilic granules of the neutrophils. Since ⁇ -defensins cause cell dysfunction such as a hemolyzing property, they are used exclusively inside the neutrophil.
  • the fourth point is mentioned that the hBDs1 to 4, ⁇ -defensins, are secreted from epithelial cells and/or keratinocytes by the action of IL-17F, which is produced from Th17-17 cells.
  • inflammatory IL-22 causes keratinocytes to secrete hBD3 therefrom to kill bacteria, but with psoriasis, IL-22 and the hBD3 are excessively secreted.
  • the MRE complex ligand which is ineffective for suppressing IL-22, is not capable of healing psoriasis, which is also consistent with clinical trials.
  • a CD4 humoral immune process is activated through a Th17 activation resulting in secretion of IgG from B cells and IgA from mucosal epithelial cells , thereby bringing on an antibacterial effects.
  • the IL-23 production test using human blood has also confirmed an average secretion 1.92 times the normal level (See Table 11).
  • the anti-viral processes with the MRE complex ligand comprise the following.
  • the first one is the process by which TLR3, TLR7, TLR9, RIG1, and MAD5, innate immune receptors, are stimulated by the MRE complex ligand or the like, thereby directly secreting IFN and the like.
  • Mainly released are IFN- ⁇ from macrophages, microglia and the like; IFN- ⁇ from epithelial cells, fibroblasts, keratinocytes, osteoblasts and the like.
  • IFN- ⁇ and IFN- ⁇ are sensed by the neighboring cells that have IFN receptors, resulting in a concurrent release of antiviral substances such as ISG or the like to exert antiviral effects.
  • IFN- ⁇ 5 The MRE complex ligand stimulation of the innate immune receptors has directly caused IFN- ⁇ 5 to be expressed at 1.6 times the normal level, IFN- ⁇ 6 1.2 times, and IFN- ⁇ 1.4-times. IFN- ⁇ serves to enhance the ability to attack virus-infected cells with a NK-cell activating interferon.
  • IFN- ⁇ is actually produced at an average of 2.89 times the normal level.
  • the second one is the production of type I IFN through IL-21.
  • the MRE complex ligand is involved in activating the non-inflammatory Th17 process, producing IL-21.
  • the IL-21 serves to activate the NK cells and also to promote production of type I IFN from epithelial cells, fibroblasts, keratinocytes, macrophages, and the like, whereby IFN- ⁇ and /or IFN- ⁇ cause antiviral substances to be released from still more cells.
  • the third one is that the MRE complex ligand produces IL-17F from Th17 and serves as an adjuvant to activate the humoral immune process, whereby in case there is a viral infection, it will play the role of enhancing antibody production against the viruses.
  • the fourth one is the clearance of virus-infected cells by the natural killer cells (NK), natural killer cells T (NKT), and cytotoxic T cells (CTL).
  • NK natural killer cells
  • NKT natural killer cells T
  • CTL cytotoxic T cells
  • the MRE complex ligand activates natural killer cells.
  • the MRE complex ligand although not as a main function thereof, activates a Th12m process, thereby elevating cell-mediated immunity and also activating a process for clearing virus-infected cells and/or cancer cells.
  • the fifth one is that inactivation of viruses by oligopeptides has been found in the 1990s, so it cannot be dismissed that the MRE complex ligand, which is more than 90% made up of oligopeptides, may involve this process.
  • the MRE complex ligand has turned out to have excellent anti-viral effects by activation of the five processes.
  • the "virus” includes human immunodeficiency virus, hepatitis C, influenza virus, human papillomavirus (HPV), virus, human herpes virus, type B hepatitis virus, and DNA- and RNA- type viruses which infect fishes or higher vertebrates.
  • HSP heat shock protein
  • koto-sugi family Taxaceae
  • quercetin quercetin
  • resveratrol that restore the apoptotic function to immortalized cancer cells
  • the immune process exerts anticancer effects by having the cytotoxic CD8 + T cells (CTL) and NKT cells, and NK cells adhere to cancer cells and/or virus-infected cells, perforating the cells with perforin, and inducing cancer cells to apoptosis by granzymes.
  • CTL cytotoxic CD8 + T cells
  • NKT cells cytotoxic CD8 + T cells
  • NK cells adhere to cancer cells and/or virus-infected cells, perforating the cells with perforin, and inducing cancer cells to apoptosis by granzymes.
  • TNF- ⁇ also activates a process that causes cytochrome C to be released from mitochondria and activates caspase-8 cells to induce cancer cells to apoptosis.
  • TNF- ⁇ is known to be a substance which is highly cytotoxic to common cells and which can also trigger the onset of HIV.
  • the cell-mediated immunity is reinforced by an IL-12 activated Th process activating CTL.
  • the CTL destroys cells infected with type C hepatitis virus and /or HIV thereby causing the onset of hepatitis and/or HIV and also developing cytotoxic autoimmune diseases.
  • the anti-cancer effect of the MRE complex ligand is executed mainly by natural killer cells (cells NK) with the anticancer effect elevated with the participation of CTL cells and NKT cells.
  • NK cells are activated by the MRE complex ligand through the below processes to exert their anti-cancer effects.
  • the first one is a process in which stimuli of innate immune receptors cause type I interferon to be produced and directly activate NK cells, a process to show effects at an early period.
  • IFN- ⁇ IFN- ⁇ (IL-28A), known to strongly activates NK, is 1.84 times the normal level, as shown in Table 17, whereby it together with IFN- ⁇ and IFN- ⁇ elevates anticancer effects.
  • CXVR4 chemokine that causes cancer cells to metastasize to the lung and/or liver is suppressed to 0.67 times the normal level, thereby suppressing cancer metastasis.
  • the second one is the anti-cancer effects due to the activation ofNK cells by producing I1-21 through a non-inflammatory Th 1 process with the MRE complex ligand.
  • the third one is a process of activating NK cells by a direct action from IL-12 on NKT cells, thereby producing IL-21.
  • the fourth one is the production of IFN ⁇ by activation of the Th1 process with IL-12. It is a process for elevating the anticancer effect by activation of CTL.
  • the MRE complex ligand is capable of activating NK cells by three processes.
  • Table 18 is for a comparison test for NK cell's ability to kill cancer cells. The comparison is made for addition of no ligand vs addition of the MRE complex ligand-containing stock solution prepared in Example 1. [Table 18] Test for ability to kill cancer cells by human NK cells.
  • Subj ect Control MRE complex ligand Composition of the instant application) 6.00% Ratio to control 0.06% 0.60% 6.00% A male 52 49.5 61.5 71.7 85.6 1.73 fold B male 31 50.5 56.9 65.1 62.4 1.24 fold C female 54 19.8 38.3 46.2 56.0 2.83 fold D female 51 10.0 22.0 17.5 20.9 2.09 fold Average 32.4 47.6 51.6 56.2 1.73 fold Rating Standard High Very high Very high Very high Very high Very high
  • the test used whole blood within 30 hours after collection. After separation of the PBMC by the Ficoll-Conray density gradient centrifugation method, the specimen was washed with RPMI (containing 10% FBS) to obtain Effector cells. This was incubated at 4.0 ⁇ 10 6 cells / ml PBMC for 24 hours under the four conditions shown below (5% CO 2 , 37°C).
  • the applied MRE-complex ligand-containing stock solution concentration was set as a base level of 6.66% (v/v) concentration assuming a 100% absorption of the MRE fermentation stock into the blood, from an approximately 100ml of a daily intake of the MRE-complex ligand-containing stock solution and a typical human blood volume of 4,500 ml.
  • Ratings for activity assessment designated: Very High for 51 or above; High for 42 to 51; Standard for 14b to 24; Very low for 14 or less.
  • the MRE complex ligand can activate NK cells by a variety of routes and can induce cancer to apoptosis at a Very High level.
  • the apoptotic debris is rapidly phagocytized by macrophages without necrotizing as would with anticancer drugs and/or radiation so that the cancer is seen to shrink by itself without causing cachexia.
  • the MRE complex ligand offers good results in cases of a dog's liver cancer by intravenous injection (See Example 8); non-Hodgkin's lymphoma cancer by ingesting (See Example 13); and a renal cancer dialysis patient (See Example 39). These results are in close agreement with those of clinical results from IL-21 administration (Patent Reference 4). The precancerous polyps of viral cancer were also observed to disappear.
  • the MRE a complex ligand, was found to have anticancer effects of reducing the cancer cells without concomitant inflammation.
  • abnormal cells such as carcinoma, sarcoma, tumor, epithelioma, leukemia, lymphoma, polyp, rigid cancer, and, malignant transformation and neoplastic cells.
  • NK cells are innate immune cells that select and eliminate only cells that deviate from “normalcy.” (Non-patent reference 5).
  • the MRE complex ligand has excellent properties, not seen in any other ligands, in that while it maintains its antibacterial, antiviral, and anticancer effects, it has anti-inflammatory and tissue repair effects.
  • the anti-inflammatory effect is also a remarkable nature of the MRE complex ligand.
  • the MRE complex ligand in accordance with the present invention, suppresses inflammation by the following processes.
  • the first one reveals that it involves a pulse-wise progression of activation and a delayed suppression with a time lag so as to activate, in pulses, a coherent immune activation process, thereby holding a balance and homeostasis as an organism.
  • Activation of NF- ⁇ B and AP-1 from the stimulation of TLR, NLR, and RLR is followed by maximization of ILIB (47.641), TNFA (21.182), and IL-8 (20.431) gene expressions, whereupon a NFKBIZ (20.011) gene expression, a "trigger" for the second stage of the process occurs vigorously and production begins the next phase products IL-6(1.405), IL-12B(1.837), IL-23A(1.784), and the like.
  • TNFAIP6 37.142
  • TNFAIP3 TNFAIP3
  • NFKBIA 8.425)
  • NFKBID 2.101
  • IKBKE 1.47
  • NFKB1 2.794
  • NF- ⁇ B NF- ⁇ B genes
  • the second one is that the MRE complex ligand suppresses the production of IL-18, a trigger of inflammatory diseases.
  • the reduced IL-18 production leads to the activation of a non-inflammatory Th17 process, non-inflammatory Th1 process, and non-inflammatoryTh2 process.
  • the non-inflammatory Th17 process and non-inflammatory Th1 process are activated by the MRE complex ligand.
  • IL-17 that activates non-inflammatory humoral immunity and IL-21 that activates NK cells are secreted from Th-17.
  • IFN- ⁇ is produced from Th1 to enhance cell-mediated immunity. While both processes retain antibacterial and antiviral potency, they act to calm inflammatory diseases including autoimmune and /or allergic diseases.
  • the third one is the calming of allergic diseases and autoimmune diseases by IL-21 production.
  • the table below shows results for an IL-21production test with human blood by the MRE complex ligand.
  • NK celll's IL-21 production test The method and procedure for NK celll's IL-21 production test are given as follows.
  • the test used whole blood within 30 hours after collection. After separation of the PBMC by the Ficoll-Conray density gradient centrifugation method, the specimen was washed with RPMI (containing 10% FBS) to obtain Effector cells. This was incubated at 4.0 ⁇ 10 6 cells / ml PBMC for 24 hours under the following conditions (5% CO 2 , 37°C). Further, the applied MRE-complex ligand-containing stock solution concentration was set as a base level of 6.66% (v/v) concentration assuming a100% absorption of the MRE fermentation stock into the blood, from an approximately 100ml of a daily intake of the MRE-complex ligand-containing stock solution and a typical human blood volume of 4,500 ml.
  • the MRE complex ligand -containing solution is found to increase the IL -21 production on average to 2.11 times the normal level; it should be noted that subjects with only a low level IL-21 production to begin with tend to have a significant increase therein.
  • IL-21 is a cytokine that enhances anticancer effects by proliferating the anti-viral component secretion and natural killer cells and it is also known to induce allergy-and/or autoimmune disease-causing T cells and B cells to apoptosis.
  • the MRE complex ligand was confirmed to really increase IL-12 substantially in the human blood in this manner, which is also clearly corroborated by clinical trials for its effects, as in Examples 15 to 20.
  • Such IL-21 production-inducing effect by the MRE complex ligand is shown to be useful in preventing and treating a variety of immune diseases, such as allergic diseases (in particular, IgE regulated allergic diseases (Type I allergic reaction-related diseases) such as asthma, hay fever, atopic dermatitis, eczema, food hypersensitivity, urticaria, allergic rhinitis , allergic conjunctivitis, and the like; autoimmune diseases (rheumatoid arthritis, Crohn's disease, systemic lupus erythematosus, scleroderma, polymyositis, polychondritis, periarteritis nodosa, ankylosing spondylitis, rheumatic fever, Sjogren's syndrome, Behcet's disease, thyroiditis , diabetes type I, dermatomyositis, chronic active hepatitis, myasthenia gravis, Grave's disease, multiple sclerosis, primary
  • GM ml macrophages
  • M2 M M2
  • the temperature is raised and the pH is shifted toward the acidic side thereby lowering bacterial and /or viral activities and at the same time creating an environment to facilitate the activities of emergency enzymes including the lysosomal enzymes. This is a process at an inflammation stage.
  • the fragments of destroyed bacteria are turned into low molecular substances by lysosomal enzymes and part thereof is sensed by the LHA receptors (MHC receptors in animals) of antigen presenting cells (APC) of macrophages, dendritic cells, vascular endothelial cells, with that information stored in memory T cells and memory B cells.
  • LHA receptors MHC receptors in animals
  • APC antigen presenting cells
  • LR, NLR, and RLR receptors of the macrophage innate immune system are sensed by LR, NLR, and RLR receptors of the macrophage innate immune system; the ligand pattern at the completion of antibacterial action is recognized, whereby the macrophages switch from the M1 to the M2 state releasing IL-10 and IL-2, and at the same time producing FGF that promotes tissue repair and activating fibroblasts, osteoblasts, and the like.
  • low molecular ligands are implied to signal the antibacterial completion pattern.
  • the MRE complex ligand is truly a product obtained by degrading bacterial cells down to low molecular products, it was possible to expect its tissue repair effects.
  • the expression of FGF2, one of the cytokines that promote tissue repair was found to be 2.525 times the normal level by a DNA kinetic analysis with a microarray.
  • the clinical trials also confirmed the MRE complex ligand's tissue repair action (See Examples 25 to 29).
  • the macrophage M1 activation also activates macrophage phagocytic activity. This serves, in the blood, lymphocytes, and tissues, to phagocytize and degrade dead cells, foreign matter, and oxidized toxins such as oxidized LDL followed by discarding the residue in the bile for internal cleansing.
  • the MRE complex ligand's M1 activation is effective for removing waste product through the activation of macrophages.
  • the MRE complex ligand has excellent properties for being simultaneously anti-inflammatory and antibacterial, antiviral, and anti-cancer. Said ligand, being low molecular in the oligo range, can be readily absorbed through the intestines and mucosa. In addition, it can also be utilized as a beverage and by topical application, or for intravenous injections after purification thereof.
  • the MRE complex ligand of the present invention can be said to possess, as its function, overall properties of suppressing lymphocytic immunity and enhancing innate immunity.
  • the immune-enhancing composition of the present invention exerts various innate immune stimulant effects as described above, on being administered to mammals, including humans, dogs and the like.
  • the method of administration may be any mode of administration regularly performed in the medical or health care field as long as the immune-enhancing composition of the present invention is administered in such a state that its function can be exerted.
  • it can be administered enterally, orally, or may be done parenterally.
  • Parenteral administration methods include intravascular administration, injection around or into the tissue, subcutaneous injection, and the like; it may also be applied directly to the skin and/or mucous membranes. Additionally permitted are intraocular administration, nasal administration, transdermal administration, and transmucosal administration.
  • the immune-enhancing composition according to the present invention can be administered in any formulation commonly used in the fields of medicine or healthcare or in the field of molecular biology, as long as such aspects permit exerting the functions of said composition.
  • the immune-enhancing composition according to the present invention can be administered as a liquid composition, wherein it may be a stock solution or a dilution.
  • Example 1 Preparation of MRE complex ligand-containing stock solution by culturing and internal sporulation of an MRE symbiotic bacterial group
  • the MRE complex ligand-containing stock solution used in the present invention is prepared by degrading its own bacterial cells of the MRE symbiotic bacteria group down to a low molecular region by blended enzyme groups of the digestive enzyme group secreted from the vegetative cells of the MRE symbiotic bacterial group with a bulk-type enzyme group homologous to the lysosomal enzyme group released along with internal sporulation.
  • the culturing of the MRE bacteria group was carried out by a common culturing method known in the art for aerobic gram-positive bacteria.
  • a 1.2m 3 culture aeration vessel was filled with 1,0001 water and was aerated.
  • the culture aeration vessel was fed with nutrients: 10kg fish meal, 10kg rice bran, 5kg oil meal, 1kg broth, along with appropriate amounts of minerals such as magnesium sulfate, silica, and the like.
  • the MRE bacterial group in the vegetative cell state along with the culture medium thereof, is separated into another aeration type culture vessel.
  • this culture aeration vessel the system is placed under a starvation condition with all the MRE bacterial group's nutrients cut off, with continued aeration under the conditions of 25 °C to 35°C, whereupon the depletion of nitrogen components triggers an internal sporulation to begin.
  • the aeration oxygen feed
  • the internal spores all begin to precipitate providing a transparent supernatant liquid.
  • the resultant supernatant liquid is pressure filtered through a 0.2 ⁇ m membrane, to obtain a low molecular MRE complex ligand-containing stock solution.
  • the MRE complex ligand-containing stock solution has 3.6 mg/ml of organic components, and contains therein 80 ⁇ g / ml of the MRE complex ligand component. It is noted in the present specification that the resultant "MRE complex ligand-containing stock solution” is called “The MRE stock solution”; the expression “dilution of the MRE complex ligand “refers to "dilution of the MRE complex ligand-containing stock solution” or “dilution of the MRE stock solution,” unless specifically stated otherwise.
  • Warts are said to be caused by HPV (human papillomavirus).
  • ALT GGT
  • a male 66 years of age. was under an anticancer treatment for pancreatic cancer. Also received insulin injections for diabetes. Ingested 30 ml of the MRE ligand beverage a day for several months; the patient is in a very good physical condition.
  • the present invention can be modified in various ways, though needless to mention, with the various modification being not limited to the embodiment described above, within the scope of not changing the gist of the invention.

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Veterinary Medicine (AREA)
  • Public Health (AREA)
  • General Health & Medical Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Animal Behavior & Ethology (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • General Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Engineering & Computer Science (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Immunology (AREA)
  • Epidemiology (AREA)
  • Mycology (AREA)
  • Microbiology (AREA)
  • Virology (AREA)
  • Molecular Biology (AREA)
  • Communicable Diseases (AREA)
  • Oncology (AREA)
  • Dermatology (AREA)
  • Diabetes (AREA)
  • Rheumatology (AREA)
  • Polymers & Plastics (AREA)
  • Physical Education & Sports Medicine (AREA)
  • Pulmonology (AREA)
  • Food Science & Technology (AREA)
  • Nutrition Science (AREA)
  • Orthopedic Medicine & Surgery (AREA)
  • Ophthalmology & Optometry (AREA)
  • Hematology (AREA)
  • Tropical Medicine & Parasitology (AREA)
  • Biotechnology (AREA)
  • Obesity (AREA)
  • Otolaryngology (AREA)
  • AIDS & HIV (AREA)
  • Heart & Thoracic Surgery (AREA)
EP10743592.7A 2009-02-20 2010-02-22 Composition immunopotentiatrice et son procédé de production Active EP2399595B1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
JP2009061956 2009-02-20
PCT/JP2010/001120 WO2010095463A1 (fr) 2009-02-20 2010-02-22 Composition immunopotentiatrice et son procédé de production

Publications (3)

Publication Number Publication Date
EP2399595A1 true EP2399595A1 (fr) 2011-12-28
EP2399595A4 EP2399595A4 (fr) 2012-11-28
EP2399595B1 EP2399595B1 (fr) 2015-11-04

Family

ID=42633750

Family Applications (1)

Application Number Title Priority Date Filing Date
EP10743592.7A Active EP2399595B1 (fr) 2009-02-20 2010-02-22 Composition immunopotentiatrice et son procédé de production

Country Status (8)

Country Link
US (1) US8454979B2 (fr)
EP (1) EP2399595B1 (fr)
JP (1) JP4951150B2 (fr)
KR (1) KR101656252B1 (fr)
CN (2) CN102387808A (fr)
AU (1) AU2010216926B2 (fr)
CA (1) CA2786891C (fr)
WO (1) WO2010095463A1 (fr)

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP3533466A4 (fr) * 2016-10-28 2020-06-10 Toray Industries, Inc. Composition pharmaceutique pour le traitement et/ou la prévention du cancer

Families Citing this family (13)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2004043405A2 (fr) * 2002-11-12 2004-05-27 The Brigham And Women's Hospital, Inc. Vaccin a base de polysaccharide destine a combattre les infections aux staphylocoques
EP2468769A1 (fr) 2004-04-21 2012-06-27 The Brigham and Women's Hospital, Inc. Peptides à liaison de glucosamine poly-N-acétyle (PNAG/DPNAG) et procédés d'utilisation
KR20110031393A (ko) 2008-07-21 2011-03-25 더 브리검 앤드 우먼즈 하스피털, 인크. 합성 베타-1,6 글루코사민 올리고당에 관한 방법 및 조성물
US9102957B2 (en) 2010-02-05 2015-08-11 Meisho Co., Ltd. Method for degrading organic material using mother cell lyases formed in association with spore formation of microorganism
JP2012136450A (ja) * 2010-12-24 2012-07-19 Nitto Yakuhin Kogyo Kk インフルエンザウイルス感染予防用組成物
AU2012227444B2 (en) * 2011-03-11 2017-03-30 Meisho.Co., Ltd Method for raising plants and composition used therefor
US9828612B2 (en) * 2011-05-23 2017-11-28 Meisho Co., Ltd. Method for producing alcohol using tree as starting material and alcohol solution obtained by same
TWI507136B (zh) * 2011-08-26 2015-11-11 Meisho Co Ltd The method of producing eggs, eggs and their use of chicken feed
FR3007655B1 (fr) * 2013-06-28 2016-12-23 Lesaffre & Cie Souche bacillus subtilis pour le traitement et/ou la prevention de maladies inflammatoires chroniques
CN108102974B (zh) * 2018-02-02 2021-07-27 山西大学 一种高产胞外多糖旧金山乳杆菌Ls-1001菌株培养方法
WO2019160116A1 (fr) * 2018-02-16 2019-08-22 イチビキ株式会社 Bactérie bacillus, agent induisant la production d'interleukines-22, et agent renforçant les fonctions de barrière cutanée
US11814506B2 (en) 2019-07-02 2023-11-14 Marathon Petroleum Company Lp Modified asphalts with enhanced rheological properties and associated methods
KR102337884B1 (ko) * 2021-07-15 2021-12-10 주식회사 여섯발자국 반려동물용 면역강화 조성물

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2003055513A2 (fr) * 2001-12-21 2003-07-10 Kings's College London Applications de spores
US20060134136A1 (en) * 2002-09-06 2006-06-22 Mcintyre Graham Whole bacterial cells as immune modulator

Family Cites Families (14)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPH0527939A (ja) * 1991-07-18 1993-02-05 Canon Inc 情報処理装置
JPH05279394A (ja) * 1992-03-02 1993-10-26 Japan Energy Corp 主要組織適合複合体クラス−ｉ遺伝子産物発現促進因子及びその製造方法
JP4074006B2 (ja) 1998-06-24 2008-04-09 アサマ化成株式会社 ＩｇＥ抗体産生抑制物質の製造方法並びにそれを用いた食品
JP2003071479A (ja) * 2001-08-31 2003-03-11 Yoji Nagahama 微生物反応装置及びそれを用いた微生物栄養源含有液の処理方法
EP1589990B1 (fr) 2002-03-27 2012-11-14 The Government of the United States of America, represented by The Secretary Department of Health and Human services Il-21 pour une utilisation dans le traitement du cancer
AU2003251418A1 (en) 2002-06-07 2003-12-22 Robert J. Chalifour Amidine derivatives for treating amyloidosis
ATE546150T1 (de) 2002-06-07 2012-03-15 Zymogenetics Inc Verwendung von il-21 und monoklonalem antikörper zur behandlung von fester tumore
GB0220809D0 (en) * 2002-09-06 2002-10-16 Univ College Of London Immune modulator
CA2538083A1 (fr) 2003-09-25 2005-04-07 Zymogenetics, Inc. Procedes de traitement de maladies auto-immunes utilisant l'interleukine-21
JP2006137703A (ja) * 2004-11-12 2006-06-01 Hakuju Life Science Co Ltd 植物免疫活性物質およびその製造方法
JP2006265212A (ja) 2005-03-25 2006-10-05 Institute Of Physical & Chemical Research Ｉｌ−２１産生誘導剤
JP4198728B2 (ja) * 2005-09-30 2008-12-17 株式会社グローバルエンジニアリング 動物用免疫活性調整剤、及びその調整剤を含有する動物用飼料
JP5019961B2 (ja) 2006-06-26 2012-09-05 株式会社ヤクルト本社 インターロイキン１０産生促進剤
JP4930717B2 (ja) 2007-09-07 2012-05-16 日本精機株式会社 車両用表示装置

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2003055513A2 (fr) * 2001-12-21 2003-07-10 Kings's College London Applications de spores
US20060134136A1 (en) * 2002-09-06 2006-06-22 Mcintyre Graham Whole bacterial cells as immune modulator

Non-Patent Citations (4)

* Cited by examiner, † Cited by third party
Title
A. TAJIMA ET AL: "Inhibition of interleukin-8 production in human endothelial cells by Staphylococcus aureus supernatant", CLINICAL & EXPERIMENTAL IMMUNOLOGY, vol. 147, no. 1, 6 December 2006 (2006-12-06), pages 148-154, XP55041056, ISSN: 0009-9104, DOI: 10.1111/j.1365-2249.2006.03254.x *
K. CHAKRABARTY ET AL: "Bacillus anthracis Spores Stimulate Cytokine and Chemokine Innate Immune Responses in Human Alveolar Macrophages through Multiple Mitogen-Activated Protein Kinase Pathways", INFECTION AND IMMUNITY, vol. 74, no. 8, 21 July 2006 (2006-07-21), pages 4430-4438, XP55041057, ISSN: 0019-9567, DOI: 10.1128/IAI.00446-06 *
MAKINO S ET AL: "Immunomodulatory effects of polysaccharidesproduced by Lactobacillus delbrueckii ssp. bulgaricus0LL1073R-1", JOURNAL OF DAIRY SCIENCE, AMERICAN DAIRY SCIENCE ASSOCIATION, US, vol. 89, no. 8, 1 August 2006 (2006-08-01) , pages 2873-2881, XP003022695, ISSN: 0022-0302, DOI: 10.3168/JDS.S0022-0302(06)72560-7 *
See also references of WO2010095463A1 *

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP3533466A4 (fr) * 2016-10-28 2020-06-10 Toray Industries, Inc. Composition pharmaceutique pour le traitement et/ou la prévention du cancer

Also Published As

Publication number Publication date
AU2010216926B2 (en) 2014-02-27
CN102387808A (zh) 2012-03-21
US20120039946A1 (en) 2012-02-16
US8454979B2 (en) 2013-06-04
EP2399595A4 (fr) 2012-11-28
EP2399595B1 (fr) 2015-11-04
JP4951150B2 (ja) 2012-06-13
CA2786891A1 (fr) 2010-08-26
CN103989713A (zh) 2014-08-20
CN103989713B (zh) 2017-12-05
KR101656252B1 (ko) 2016-09-09
AU2010216926A1 (en) 2011-10-13
WO2010095463A1 (fr) 2010-08-26
KR20110118724A (ko) 2011-10-31
JPWO2010095463A1 (ja) 2012-08-23
CA2786891C (fr) 2017-08-01

Similar Documents

Publication Publication Date Title
US8454979B2 (en) Immunopotentiating composition and process for producing same
Legrand et al. Lactoferrin: Lactoferrin: a modulator of immune and inflammatory responses
Kohchi et al. Applications of lipopolysaccharide derived from Pantoea agglomerans (IP-PA1) for health care based on macrophage network theory
CA3004951A1 (fr) Regulation des niveaux d'oxydoreduction cellulaire
NZ261094A (en) Manufacture of a medicament for stimulating a human immune system involving administration of a mycobacterium extract that does not contain oil
US9056073B2 (en) Vaccine comprising lactobacilli for treating prostate inflammation and benign prostate hyperplasias
US20040208896A1 (en) Immunotherapy for humans
Jain et al. Immunostimulants: concepts, types and functions
WO2005077390A1 (fr) Agent hypoglycémiant, remède/préventif pour le diabète et leur procédé de fabrication
CN115216423B (zh) 一种动物双歧杆菌sf及其在医药和食品中的应用
Kusumawardani et al. The effect of ethanolic extract of propolis on skin manifestation and skin tissue necrosis in cutaneous anthrax animal model
JPH09263539A (ja) 抗真菌作用を有する皮膚疾患治療剤
EP2486931B1 (fr) Agent thérapeutique pour des maladies infectieuses causées par le virus de la grippe
JPH1192389A (ja) 免疫賦活剤
US6806253B2 (en) Immunodulatory complex and use thereof in helicobacter diseases
KR20020021375A (ko) 미코박테리움 바카이를 활용하여 면역적으로 매개된질병을 치료하는 방법 및 조성물
CN101791318B (zh) 细菌脂多糖的新用途
JP2003155249A (ja) ＩｇＡ産生促進剤
WO2009086695A1 (fr) Utilisation d'un extrait de polysaccharides du bcg et d'acide nucléique pour préparer un médicament pour traiter une dermatose allergique, et procédé d'injection et de préparation d'un tel extrait
JP2001231495A (ja) 好酸球集積抑制作用を有する食品
Ayub Immunomodulatory Effect of Selected Fruit Waste by Product Leads on Cyclophosphamide Induced Immunosuppression in Mice
CN115747097A (zh) 一种Bifidobacterium bifidum菌株及其在肿瘤辅助治疗中的应用
RU2468816C1 (ru) Пробиотический бактериальный препарат корпускулярных антигенов коринебактерий липопептидополисахаридной природы для профилактики и лечения туберкулеза, способ его получения
CN101780271A (zh) 内毒素和凝集素的药用组合物及其用途
JP2005145831A (ja) アレルギー体質改善剤

Legal Events

Date Code Title Description
PUAI Public reference made under article 153(3) epc to a published international application that has entered the european phase

Free format text: ORIGINAL CODE: 0009012

17P Request for examination filed

Effective date: 20110919

AK Designated contracting states

Kind code of ref document: A1

Designated state(s): AT BE BG CH CY CZ DE DK EE ES FI FR GB GR HR HU IE IS IT LI LT LU LV MC MK MT NL NO PL PT RO SE SI SK SM TR

DAX Request for extension of the european patent (deleted)
A4 Supplementary search report drawn up and despatched

Effective date: 20121025

RIC1 Information provided on ipc code assigned before grant

Ipc: A61K 35/74 20060101AFI20121019BHEP

Ipc: A61K 39/07 20060101ALI20121019BHEP

Ipc: A61K 39/02 20060101ALI20121019BHEP

17Q First examination report despatched

Effective date: 20131105

GRAP Despatch of communication of intention to grant a patent

Free format text: ORIGINAL CODE: EPIDOSNIGR1

INTG Intention to grant announced

Effective date: 20150722

GRAS Grant fee paid

Free format text: ORIGINAL CODE: EPIDOSNIGR3

GRAA (expected) grant

Free format text: ORIGINAL CODE: 0009210

AK Designated contracting states

Kind code of ref document: B1

Designated state(s): AT BE BG CH CY CZ DE DK EE ES FI FR GB GR HR HU IE IS IT LI LT LU LV MC MK MT NL NO PL PT RO SE SI SK SM TR

REG Reference to a national code

Ref country code: GB

Ref legal event code: FG4D

REG Reference to a national code

Ref country code: CH

Ref legal event code: EP

REG Reference to a national code

Ref country code: AT

Ref legal event code: REF

Ref document number: 758719

Country of ref document: AT

Kind code of ref document: T

Effective date: 20151115

REG Reference to a national code

Ref country code: IE

Ref legal event code: FG4D

REG Reference to a national code

Ref country code: DE

Ref legal event code: R096

Ref document number: 602010028792

Country of ref document: DE

REG Reference to a national code

Ref country code: FR

Ref legal event code: PLFP

Year of fee payment: 7

REG Reference to a national code

Ref country code: NL

Ref legal event code: MP

Effective date: 20151104

REG Reference to a national code

Ref country code: LT

Ref legal event code: MG4D

REG Reference to a national code

Ref country code: AT

Ref legal event code: MK05

Ref document number: 758719

Country of ref document: AT

Kind code of ref document: T

Effective date: 20151104

PG25 Lapsed in a contracting state [announced via postgrant information from national office to epo]

Ref country code: NO

Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT

Effective date: 20160204

Ref country code: HR

Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT

Effective date: 20151104

Ref country code: IS

Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT

Effective date: 20160304

Ref country code: LT

Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT

Effective date: 20151104

Ref country code: NL

Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT

Effective date: 20151104

Ref country code: ES

Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT

Effective date: 20151104

PG25 Lapsed in a contracting state [announced via postgrant information from national office to epo]

Ref country code: AT

Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT

Effective date: 20151104

Ref country code: BE

Free format text: LAPSE BECAUSE OF NON-PAYMENT OF DUE FEES

Effective date: 20160229

Ref country code: GR

Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT

Effective date: 20160205

Ref country code: LV

Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT

Effective date: 20151104

Ref country code: FI

Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT

Effective date: 20151104

Ref country code: PL

Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT

Effective date: 20151104

Ref country code: PT

Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT

Effective date: 20160304

Ref country code: SE

Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT

Effective date: 20151104

PG25 Lapsed in a contracting state [announced via postgrant information from national office to epo]

Ref country code: CZ

Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT

Effective date: 20151104

REG Reference to a national code

Ref country code: DE

Ref legal event code: R097

Ref document number: 602010028792

Country of ref document: DE

PG25 Lapsed in a contracting state [announced via postgrant information from national office to epo]

Ref country code: RO

Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT

Effective date: 20151104

Ref country code: DK

Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT

Effective date: 20151104

Ref country code: EE

Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT

Effective date: 20151104

Ref country code: SK

Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT

Effective date: 20151104

Ref country code: SM

Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT

Effective date: 20151104

PLBE No opposition filed within time limit

Free format text: ORIGINAL CODE: 0009261

STAA Information on the status of an ep patent application or granted ep patent

Free format text: STATUS: NO OPPOSITION FILED WITHIN TIME LIMIT

PG25 Lapsed in a contracting state [announced via postgrant information from national office to epo]

Ref country code: MC

Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT

Effective date: 20151104

Ref country code: LU

Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT

Effective date: 20160222

REG Reference to a national code

Ref country code: CH

Ref legal event code: PL

26N No opposition filed

Effective date: 20160805

PG25 Lapsed in a contracting state [announced via postgrant information from national office to epo]

Ref country code: CH

Free format text: LAPSE BECAUSE OF NON-PAYMENT OF DUE FEES

Effective date: 20160229

Ref country code: LI

Free format text: LAPSE BECAUSE OF NON-PAYMENT OF DUE FEES

Effective date: 20160229

PG25 Lapsed in a contracting state [announced via postgrant information from national office to epo]

Ref country code: SI

Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT

Effective date: 20151104

REG Reference to a national code

Ref country code: IE

Ref legal event code: MM4A

PG25 Lapsed in a contracting state [announced via postgrant information from national office to epo]

Ref country code: BE

Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT

Effective date: 20151104

PG25 Lapsed in a contracting state [announced via postgrant information from national office to epo]

Ref country code: IE

Free format text: LAPSE BECAUSE OF NON-PAYMENT OF DUE FEES

Effective date: 20160222

REG Reference to a national code

Ref country code: FR

Ref legal event code: PLFP

Year of fee payment: 8

PG25 Lapsed in a contracting state [announced via postgrant information from national office to epo]

Ref country code: MT

Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT

Effective date: 20151104

REG Reference to a national code

Ref country code: FR

Ref legal event code: PLFP

Year of fee payment: 9

PG25 Lapsed in a contracting state [announced via postgrant information from national office to epo]

Ref country code: CY

Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT

Effective date: 20151104

Ref country code: HU

Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT; INVALID AB INITIO

Effective date: 20100222

PG25 Lapsed in a contracting state [announced via postgrant information from national office to epo]

Ref country code: MT

Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT

Effective date: 20160229

Ref country code: TR

Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT

Effective date: 20151104

Ref country code: MK

Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT

Effective date: 20151104

PG25 Lapsed in a contracting state [announced via postgrant information from national office to epo]

Ref country code: BG

Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT

Effective date: 20151104

PGFP Annual fee paid to national office [announced via postgrant information from national office to epo]

Ref country code: FR

Payment date: 20230210

Year of fee payment: 14

PGFP Annual fee paid to national office [announced via postgrant information from national office to epo]

Ref country code: IT

Payment date: 20230203

Year of fee payment: 14

PGFP Annual fee paid to national office [announced via postgrant information from national office to epo]

Ref country code: DE

Payment date: 20240214

Year of fee payment: 15

Ref country code: GB

Payment date: 20240125

Year of fee payment: 15